K107R induces gene expression in an adipocyte-autonomous manner. K107R induces gene expression in an adipocyte-autonomous manner. (A) Comparison of gene regulation by K107R and rosiglitazone in vivo and in vitro. For the in vivo analysis, RNA was prepared from iWAT and eWAT from WT and K107R mice administered rosiglitazone or vehicle by oral gavage for 7 d. For the in vitro analysis, RNA was prepared from WT and K107R stromal vascular cells derived from iWAT and differentiated for 8 d using the insulin/dexamethasone/IBMX mixture and subsequently incubated for 48 h with either rosiglitazone or vehicle. Gene expression was measured by qPCR. Cycle threshold (Ct) values are shown for WT/vehicle conditions. (B) Immunoblotting for SUMO1-conjugated PPARγ (SUMO1) and total PPARγ following PPARγ immunoprecipitation (IP) (Top Two Rows) and total PPARγ, UBC9, and β-actin in total cell extract (Bottom Three Rows) from WT and K107R adipocytes treated with a control or UBC9 siRNA. (C) Quantification of Fgf21 and Elovl3 gene expression by qPCR using RNA prepared from WT and K107R adipocytes exposed to either the UBC9 or control siRNA. Ct values are shown for WT/control siRNA conditions. For B and C, WT and K107R stromal vascular cells were treated with control or UBC9 siRNA for 2 d in growth medium. The cells were subsequently incubated for 2 d in differentiation medium containing insulin, dexamethasone, IBMX, and rosiglitazone, followed by 2 d in medium containing only insulin and rosiglitazone. *P < 0.05; **P < 0.01; ***P < 0.001. Takeshi Katafuchi et al. PNAS 2018;115:48:12102-12111 ©2018 by National Academy of Sciences